MODIFIED MRNAS ENCODING CELL-PENETRATING POLYPEPTIDES
This invention relates to modified nucleic acid compositions encoding cell-penetrating polypeptides to provoke an innate immune response in a cell and methods of delivering protein-binding partners to target cells.
1 . A composition comprising a modified nucleic acid encoding a cell-penetrating polypeptide, wherein the modified nucleic acid provokes a reduced innate immune response as compared to an unmodified nucleic acid encoding the same cell-penetrating polypeptide when introduced into a first cell.
2 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a complex comprising a charged protein operably linked to a therapeutic protein.
3 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a fusion protein comprising a charged protein covalently linked to a therapeutic protein.
4 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a therapeutic protein comprising a surface charge.
5 . The composition of claim 1 , wherein the cell-penetrating polypeptide further comprises a detectable label.
6 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a supercharged polypeptide.
7 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a signal sequence for secretion of the cell-penetrating polypeptide.
8 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a ratio of surface charge to surface amino acids is approximately 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 or 0.9.
9 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a first domain comprising a supercharged polypeptide and a second domain comprising a protein-binding partner.
10 . The composition of claim 9 , wherein the cell-penetrating polypeptide is capable of penetrating a second cell.
11 . The composition of claim 10 , wherein the first cell and the second cell are cells from the same tissue.
12 . The composition of claim 10 , wherein the first cell and the second cell are cells from different tissues.
13 . The composition of claim 10 , wherein the first cell is located proximal to the second cell.
14 . The composition of claim 10 , wherein the first cell is located distal to the second cell.
15 . The composition of claim 10 , wherein the cell-penetrating polypeptide penetrating the second cell comprises an intracellular binding partner for the protein-binding partner.
16 . The composition of claim 9 , wherein the cell-penetrating polypeptide is capable of penetrating the first cell.
17 . The composition of claim 1 , wherein the cell-penetrating polypeptide comprises a protein-binding partner.
18 . The composition of claim 17 , wherein the protein-binding partner is selected from a supercharged antibody and a functional fragment thereof.
19 . The composition of claim 17 where the modified nucleic acid is introduced to a cell where the cell-penetrating polypeptide is secreted from.
20 . The composition of claim 19 , wherein the cell-penetrating polypeptide is capable of penetrating the second cell comprising an intracellular binding partner for the supercharged antibody or functional fragment thereof.
21 . The composition of claim 19 , wherein the cell-penetrating polypeptide is capable of penetrating the first cell.
22 . A formulation comprising the composition of claim 1 and a delivery agent.
23 . The formulation of claim 22 , wherein the delivery agent comprises a cationic material.
24 . The formulation of claim 22 , wherein the delivery agent comprises a cell-penetrating polypeptide.
25 . A method of delivering a protein-binding partner into a target cell comprising an intracellular binding partner for the protein-binding partner, comprising the step of introducing into a first cell a composition comprising a modified nucleic acid encoding a cell-penetrating polypeptide comprising the protein-binding partner under conditions wherein the cell-penetrating polypeptide is produced in and secreted from the first cell, and subsequently penetrates into the target cell.
26 . The method of claim 25 , wherein the modified nucleic acid provokes a reduced innate immune response as compared to an unmodified nucleic acid encoding the same cell-penetrating polypeptide when introduced into the first cell.
27 . The method of claim 25 , wherein the first cell and the target cell are the same cell.
28 . The method of claim 25 , wherein the first cell and the target cell are co-located in a tissue of a subject.
29 . The method of claim 25 , wherein the first cell is located proximal to the target cell.
30 . The method of claim 25 , wherein the first cell is located distal to the target cell.
31 . The method of claim 25 , wherein the modified nucleic acid is retained in an extracellular depot adjacent to the first cell prior to introduction in the first cell.